CEN ISO/TS 6579-2:2012
(Main)Microbiology of food and animal feed - Horizontal method for the detection, enumeration and serotyping of Salmonella - Part 2: Enumeration by a miniaturized most probable number technique (ISO/TS 6579-2:2012)
Microbiology of food and animal feed - Horizontal method for the detection, enumeration and serotyping of Salmonella - Part 2: Enumeration by a miniaturized most probable number technique (ISO/TS 6579-2:2012)
ISO/TS 6579-2 gives a method for the enumeration of Salmonella spp. present in: products intended for human consumption and for the feeding of animals; environmental samples in the area of food production and food handling; animal faeces; and environmental samples from the primary production stage by calculation of the most probable number (MPN).
The method is based on miniaturization of the dilution, pre-enrichment and selective enrichment steps. The selective enrichment medium, modified semi-solid Rappaport?Vassiliadis (MSRV), is intended for the detection of motile salmonellae and is not appropriate for the detection of non-motile salmonellae.
It is possible that the method is less appropriate to enumerate Salmonella ser. Typhi and Salmonella ser. Paratyphi.
The method is not appropriate for the enumeration of Salmonella spp. in (very) low contaminated samples (<1 cfu/g).
Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren zum Nachweis, zur Zählung und zur Serotypisierung von Salmonellen - Teil 2: Zählung unter Anwendung eines miniaturisierten Verfahrens der wahrscheinlichsten Keimzahl (ISO/TS 6579-2:2012)
Dieser Teil von ISO 6579 legt ein Verfahren zur Zählung von Salmonella spp. in
- Produkten, die für den menschlichen Verzehr oder als Futtermittel bestimmt sind;
- Umgebungsproben im Bereich der Herstellung und Handhabung von Lebensmitteln;
- Tierkot;
- Umgebungsproben im Bereich der Primärproduktion;
fest, und zwar durch Berechnung der wahrscheinlichsten Keimzahl (en: most probable number, MPN).
Das Verfahren beruht auf einer Miniaturisierung der Schritte der Verdünnung, Voranreicherung und selektiven Anreicherung. Das selektive Anreicherungsmedium, modifiziertes halbfestes Medium nach Rappaport-Vassiliadis (MSRV), ist für den Nachweis von beweglichen Salmonellen vorgesehen. Es ist nicht für den Nachweis von unbeweglichen Salmonellen geeignet.
Es ist möglich, dass dieses Verfahren weniger für Salmonella ser. Typhi und Samonella ser. Paratyphi geeignet ist.
Das Verfahren ist nicht für die Zählung von Salmonella spp. in (sehr) gering kontaminierten Proben ( 1 KBE/g) geeignet.
ANMERKUNG Bei den meisten der für dieses Verfahren relevanten Matrices ist im Allgemeinen die Anzahl der unbeweglichen Salmonellen gering. In dieser Anmerkung sind Beispiele für Proben aus der Primärproduktion genannt. Die unbeweglichen Salmonella-Biovaren von Salmonella Gallinarum (Salmonella Gallinarum biovar gallinarum und Salmonella Gallinarum biovar pullorum) scheinen in Umgebungsproben nicht lange zu überleben und werden deshalb in Kot- oder Umgebungsproben (wie z. B. Staubproben) selten nachgewiesen (unabhängig vom Verfahren). Die Anzahl von anderen unbeweglichen Salmonella-Serovare in Kotproben scheint gewöhnlich gering zu sein. Zum Beispiel waren in einer Studie [4], in der etwa 1 000 Kotproben aus Legehennenbeständen und etwa 900 Kotproben aus Jungmastgeflügelbeständen analysiert wurden, weniger als 1 % der Gesamtanzahl der Proben in einer selektiven Bouillon positiv und auf MSRV-Medium gleichzeitig negativ (und vermutlich unbeweglich). Ähnliche Ergebnisse wurden in einer niederländischen Studie mit etwa 3 200 Kotproben von Schweinen erhalten (unveröffentlichte Daten). Andererseits wären im Fall der Studie in Literaturhinweis [4] fast 40 % der positiven Proben nicht nachgewiesen worden (d. h. falsch-negative Proben), wenn nur eine selektive Bouillon (in diesem Fall nach Rappaport-Vassiliadis) anstelle eines halbfesten Mediums verwendet worden wäre.
Microbiologie des aliments - Méthode horizontale pour la recherche, le dénombrement et le sérotypage des salmonella - Partie 2: Dénombrement par une technique miniaturisée du nombre le plus probable (ISO/TS 6579-2:2012)
L'ISO/TS 6579-2:2012 fournit une méthode de dénombrement de Salmonella spp. présentes dans: les produits destinés à la consommation humaine ou à l'alimentation animale; les échantillons environnementaux dans le domaine de la production et de la manutention de produits alimentaires; les matières fécales des animaux; les échantillons environnementaux au stade de la production primaire; par le calcul du nombre le plus probable (NPP).
La méthode est fondée sur une miniaturisation des étapes de dilution, de préenrichissement et d'enrichissement sélectif. Le milieu d'enrichissement sélectif, le milieu semi-solide de Rappaport-Vassiliadis modifié (MSRV), est conçu pour la recherche des Salmonella mobiles et n'est pas adapté à la recherche des Salmonella immobiles.
Il est possible que la méthode soit moins appropriée au dénombrement de Salmonella ser. Typhi et Salmonella ser. Paratyphi.
La méthode n'est pas appropriée au dénombrement de Salmonella spp. présentes en (très) faibles concentrations dans des échantillons contaminés (< 1 ufc/g).
Mikrobiologija živil in krme - Horizontalna metoda za ugotavljanje prisotnosti, števila in serotipov Salmonella - 2. del: Ugotavljanje števila z miniaturizirano metodo najbolj verjetnega števila (MPN) (ISO/TS 6579-2:2012)
Ta del standarda ISO 6579 podaja metodo za ugotavljanje števila Salmonella spp., prisotnih v:
– izdelkih, namenjenih za prehrano ljudi in krmo živali;
– okoljskih vzorcih na območju proizvodnje hrane in ravnanja s hrano;
– iztrebkih živali;
– okoljskih vzorcih iz faze primarne proizvodnje; z izračunom najbolj verjetnega števila (MPN). Metoda temelji na miniaturizaciji korakov redčenja, predbogatenja in selektivnega bogatenja. Medij selektivnega bogatenja, spremenjen poltrd Rappaport–Vassiliadis (MSRV), je namenjen za odkrivanje gibljivih salmonel in ni primeren za odkrivanje negibljivih salmonel. Možno je, da je metoda manj primerna za ugotavljanje števila Salmonella ser. Typhi in Salmonella ser. Paratyphi. Ta metoda ni primerna za ugotavljanje števila Salmonella spp. v (zelo) malo kontaminiranih vzorcih (< 1 cfu/g).
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
01-maj-2013
1DGRPHãþD
SIST EN ISO 6579:2003
SIST EN ISO 6579:2003/A1:2007
SIST EN ISO 6579:2003/AC:2004
SIST EN ISO 6579:2003/AC:2006
Mikrobiologija živil in krme - Horizontalna metoda za ugotavljanje prisotnosti,
števila in serotipov Salmonella - 2. del: Ugotavljanje števila z miniaturizirano
metodo najbolj verjetnega števila (MPN) (ISO/TS 6579-2:2012)
Microbiology of food and animal feed - Horizontal method for the detection, enumeration
and serotyping of Salmonella - Part 2: Enumeration by a miniaturized most probable
number technique (ISO/TS 6579-2:2012)
Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren zum
Nachweis, zur Zählung und zur Serotypisierung von Salmonellen - Teil 2: Zählung unter
Anwendung eines miniaturisierten Verfahrens der wahrscheinlichsten Keimzahl (ISO/TS
6579-2:2012)
Microbiologie des aliments - Méthode horizontale pour la recherche, le dénombrement et
le sérotypage des salmonella - Partie 2: Dénombrement par une technique miniaturisée
du nombre le plus probable (ISO/TS 6579-2:2012)
Ta slovenski standard je istoveten z: CEN ISO/TS 6579-2:2012
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
TECHNICAL SPECIFICATION
CEN ISO/TS 6579-2
SPÉCIFICATION TECHNIQUE
TECHNISCHE SPEZIFIKATION
November 2012
ICS 07.100.30
English Version
Microbiology of food and animal feed - Horizontal method for the
detection, enumeration and serotyping of Salmonella - Part 2:
Enumeration by a miniaturized most probable number technique
(ISO/TS 6579-2:2012)
Microbiologie des aliments - Méthode horizontale pour la Mikrobiologie von Lebensmitteln und Futtermitteln -
recherche, le dénombrement et le sérotypage des Horizontales Verfahren zum Nachweis, zur Zählung und zur
salmonella - Partie 2: Dénombrement par une technique Serotypisierung von Salmonellen - Teil 2: Zählung unter
miniaturisée du nombre le plus probable (ISO/TS 6579- Anwendung eines miniaturisierten Verfahrens der
2:2012) wahrscheinlichsten Keimzahl (ISO/TS 6579-2:2012)
This Technical Specification (CEN/TS) was approved by CEN on 16 July 2012 for provisional application.
The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to submit their
comments, particularly on the question whether the CEN/TS can be converted into a European Standard.
CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS available
promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in parallel to the CEN/TS)
until the final decision about the possible conversion of the CEN/TS into an EN is reached.
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© 2012 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN ISO/TS 6579-2:2012: E
worldwide for CEN national Members.
Contents Page
Foreword .3
Foreword
This document (CEN ISO/TS 6579-2:2012) has been prepared by Technical Committee ISO/TC 34 "Food
products" in collaboration with Technical Committee CEN/TC 275 “Food analysis - Horizontal methods” the
secretariat of which is held by DIN.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN/CENELEC Internal Regulations, the national standards organisations of the following
countries are bound to announce this Technical Specification: Austria, Belgium, Bulgaria, Croatia, Cyprus,
Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany,
Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO/TS 6579-2:2012 has been approved by CEN as a CEN/TS ISO 6579-2:2012 without any
modification.
TECHNICAL ISO/TS
SPECIFICATION 6579-2
First edition
2012-11-01
Microbiology of food and animal feed —
Horizontal method for the detection,
enumeration and serotyping of
Salmonella —
Part 2:
Enumeration by a miniaturized most
probable number technique
Microbiologie des aliments — Méthode horizontale pour la recherche, le
dénombrement et le sérotypage des Salmonella —
Partie 2: Dénombrement par une technique miniaturisée du nombre le
plus probable
Reference number
ISO/TS 6579-2:2012(E)
©
ISO 2012
ISO/TS 6579-2:2012(E)
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
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ii © ISO 2012 – All rights reserved
ISO/TS 6579-2:2012(E)
Contents Page
Foreword .iv
Introduction . v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Principle . 2
4.1 General . 2
4.2 Pre-enrichment in non-selective liquid medium . 2
4.3 Enrichment on a selective semi-solid medium . 2
4.4 Selective plating and identification . 2
4.5 Confirmation . 2
4.6 Calculation of most probable number . 3
5 Culture media and sera . 3
5.1 General . 3
5.2 Culture media . 3
6 Apparatus and glassware . 3
7 Sampling . 4
8 Preparation of test sample . 4
9 Procedure . 4
9.1 Test portion, initial suspension . 4
9.2 Dilution and pre-enrichment in non-selective liquid medium . 5
9.3 Selective enrichment on a semi-solid medium . 5
9.4 Selective plating . 5
9.5 Biochemical and serological confirmation . 6
10 Expression of results . 8
11 Test report . 8
Annex A (informative) Composition and preparation of culture media and reagents . 9
Annex B (informative) Diagram of procedure .17
Bibliography .18
ISO/TS 6579-2:2012(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
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The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
In other circumstances, particularly when there is an urgent market requirement for such documents, a technical
committee may decide to publish other types of document:
— an ISO Publicly Available Specification (ISO/PAS) represents an agreement between technical experts in
an ISO working group and is accepted for publication if it is approved by more than 50 % of the members
of the parent committee casting a vote;
— an ISO Technical Specification (ISO/TS) represents an agreement between the members of a technical
committee and is accepted for publication if it is approved by 2/3 of the members of the committee
casting a vote.
An ISO/PAS or ISO/TS is reviewed after three years in order to decide whether it will be confirmed for a further
three years, revised to become an International Standard, or withdrawn. If the ISO/PAS or ISO/TS is confirmed,
it is reviewed again after a further three years, at which time it must either be transformed into an International
Standard or be withdrawn.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO/TS 6579-2 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 9,
Microbiology.
ISO 6579 consists of the following part, under the general title Microbiology of food and animal feed —
Horizontal method for the detection, enumeration and serotyping of Salmonella:
— Part 2: Enumeration·by·a·miniaturized·most·probable·number·technique [Technical Specification]
Additional parts, dealing with a detection method and guidance for serotyping are planned. ISO 6579:2002 is
to be withdrawn at a later date.
iv © ISO 2012 – All rights reserved
ISO/TS 6579-2:2012(E)
Introduction
The procedure described is based on the method reported in Reference [1].
The enumeration procedure described here concerns a miniaturized most probable number (MPN) technique.
For this mini-MSRV (modified semi-solid Rappaport–Vassiliadis) MPN technique, the volume of
...
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